Geometrical Micropillars Combined with Chemical Surface Modifications – Independency of Actin Filament Spatial Distribution in Primary Osteoblasts

Abstract

Cell-biomaterial interactions are strongly affected by topographical and chemical surface characteristics. We found out earlier that geometric titanium (Ti) pillar structures in the micrometer range induce the cells to rearrange their actin cytoskeleton in short fibers solely on the top of the pillars. As a result, cell physiology was hampered concerning collagen I synthesis and spreading capacity. Furthermore, the position-dependent initial cell adhesion strength was declined near the edges. We asked whether these observed cellular effects can be performed only in combination with Ti or occur independently of chemical surface features. In addition, the specific culture conditions, e.g. serum content or influence of gravity, were of interest. Human primary osteoblasts were cultured in Osteoblast Growth Medium with serum containing SupplementMix on pure silicon pillars (5x5x5 μm) or on samples additionally sputtered with Ti (as reference) or gold. To offer the cells ligands for their adhesion receptors, we coated the pillars with collagen I or alternatively with a plasma polymer layer from allylamine. Different from standard culture conditions, the cells were cultured against gravity as well as without serum. The actin cytoskeleton was stained with phalloidin-TRITC after 24 h and analyzed by confocal laser scanning microscopy. Interestingly, on all modifications tested the cell’s actin cytoskeleton was distinctly organized in short fibers on the top of the pillars. Thus, we were able to exclude the influence of (i) the material chemistry (gold, silicon, physical plasma vs. Ti), (ii) the protein deposition on the pillar top and edges, and (iii) the impression caused by gravity.